Bacterial pathogens in rainbow trout, Oncorhynchus mykiss (Walbaum), reared at Danish freshwater farms

During a 2-year period, bacterial fish pathogens were monitored on five rainbow trout, Oncorhynchus mykiss (Walbaum), freshwater farms in Denmark. A total of 1206 fish were examined and 361 bacterial isolates were identified phenotypically. Enteric redmouth disease, furunculosis and rainbow trout fry syndrome/coldwater disease were recorded. Infections caused by Flavobacterium psychrophilum occurred most frequently, but only one outbreak of enteric redmouth disease caused by Yersinia ruckeri serotype O1 and one of furunculosis caused by Aeromonas salmonicida were recorded during the monitoring period. Flavobacterium psychrophilum was isolated on all farms, both during disease outbreaks and from fish without any signs of disease. Serological investigations of F. psychrophilum showed that serotype Th was the dominant serotype found. The serotypes Th and Fd were involved in disease outbreaks of fry and larger fish. All isolates of F. psychrophilum showed proteolytic activities; however, a few isolates, belonging to serotype FpT did not degrade elastin and were not associated with mortality. Increasing resistance problems to oxytetracycline were demonstrated. More than half of the F. psychrophilum isolates showed resistance to oxolinic acid and oxytetracycline. No antibiotic resistant isolates were found among Y. ruckeri and A. salmonicida .     

Susceptibility of Chinook salmon,Oncorhynchus tshawytscha (Walbaum), and rainbow trout,Salmo gairdneri Richardson,to infection with Vibrio anguillarum following sublethal copper exposure*

Abstract. Chinook salmon, Oncorhynchus tshawytscha (Walbaum), and rainbow trout, Salmo gairdneri Richardson, were used to determine if sublethal copper exposure would increase their susceptibility to Vibrio anguillarum infection. Fish were pretreated with copper at fractional levels of the 96 h copper LC₅₀ before exposure to the pathogen. Mortality by vibriosis was greater among fish exposed to 9% of the copper LC₅₀ for 96 h than unexposed fish. Peak susceptibility to vibriosis depended in part on the interaction of exposure time and copper concentration. The higher copper concentrations produced peak susceptibility to infection in shorter time periods. After the peak of susceptibility, sensitivity to infection declined to near control levels in those fish where exposure was continued. Rainbow trout stressed by copper required about 50% fewer pathogens to induce a fatal infection than non-copper exposed fish.     

Cellular components of probiotics control Yersinia ruckeri infection in rainbow trout, Oncorhynchus mykiss (Walbaum)

Subcellular components of the probiotics Aeromonas sobria GC2 and Bacillus subtilis JB-1, when administered to rainbow trout, Oncorhynchus mykiss , conferred protection against a new biogroup of Yersinia ruckeri . Thus, intraperitoneal or intramuscular injection of rainbow trout with cell wall proteins (CWPs), outer membrane proteins (OMPs), lipopolysaccharides (LPS), whole cell proteins (WCPs) and live cells followed by challenge on day 8 with Y. ruckeri led to 80–100% survival compared with 10% survival in the controls. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles of WCPs and OMPs from GC2 had 10 and 5 variable protein bands in comparison to 11 and 5 bands in the WCPs and CWPs from JB-1. Proteomic analyses were employed following SDS-PAGE to categorize one dominant protein of 104.7 kDa from the CWPs of JB-1 and equated it with ' Bacillus spp. endoglucanase' with a Mascot score >69. These results point to the potential of using cellular components of probiotics for protection of fish against bacterial diseases.     

Susceptibility of steelhead trout Salmo gairdneri Richardson to redmouth infection Yersinia ruckeri following exposure to copper

Abstract. Steelhead trout were exposed to sublethal concentrations of copper and inoculated with Yersinia ruckeri. Copper exposure at 7 and 10 μg/1 for 96 h caused more fish to die of infection than control fish (no copper). Infection susceptibility increased with time of exposure to a single dosage of copper (10 μg/1), reaching a maximum at 48 h. Lowering the copper concentration to 5 μg/1 caused the infection susceptibility to occur at 24 h. The infectious dose of Y. ruckeri was lower in fish exposed to 10 μg/1 copper for 48 h than control fish.     

Relative resistance of Pacific salmon to infectious salmon anaemia virus

Infectious salmon anaemia (ISA) is a major disease of Atlantic salmon, Salmo salar, caused by an orthomyxovirus (ISAV). Increases in global aquaculture and the international movement of fish made it important to determine if Pacific salmon are at risk. Steelhead trout, Oncorhynchus mykiss, and chum, O. keta, Chinook, O. tshawytscha, coho, O. kisutch, and Atlantic salmon were injected intraperitoneally with a high, medium, or low dose of a Norwegian strain of ISAV. In a second challenge, the same species, except chum salmon, were injected with a high dose of either a Canadian or the Norwegian strain. Average cumulative mortality of Atlantic salmon in trial 1 was 12% in the high dose group, 20% in the medium dose group and 16% in the low dose group. The average cumulative mortality of Atlantic salmon in trial 2 was 98%. No signs typical of ISA and no ISAV-related mortality occurred among any of the groups of Oncorhynchus spp. in either experiment, although ISAV was reisolated from some fish sampled at intervals post-challenge. The results indicate that while Oncorhynchus spp. are quite resistant to ISAV relative to Atlantic salmon, the potential for ISAV to adapt to Oncorhynchus spp. should not be ignored.     

Genetic variation in mortality of chinook salmon, Oncorhynchus tshawytscha (Walbaum), challenged with high water temperatures

Abstract. Chinook salmon, Oncorhynchus tshawytscha (Walbaum), from a northern streamtype (Bear River) and a southern ocean-type (Robertson Creek) population in British Columbia were mated within populations in a nested breeding design and the resulting juveniles were challenged with high water temperatures. Juveniles in the southern population died significantly later than those in the northern population, but none of the variation among families in mortality rate or time to death in the southern population was attributable to additive genetic variation. Non-additive genetic or maternal effects or both were observed for mortality rates in the Robertson Creek population, whereas additive genetic effects were observed for mortality at temperatures less than 22°C in the Bear River population. Larger fish survived better during the challenges than did smaller fish, and there was a negative genetic correlation (based on dam variance components) between body weight and mortality.     

Effects of temperature on Renibacterium salmoninarum infection and transmission potential in Chinook salmon,Oncorhynchus tshawytscha (Walbaum)

Renibacterium salmoninarum is a significant pathogen of salmonids and the causative agent of bacterial kidney disease (BKD). Water temperature affects the replication rate of pathogens and the function of the fish immune system to influence the progression of disease. In addition, rapid shifts in temperature may serve as stressors that reduce host resistance. This study evaluated the effect of shifts in water temperature on established R. salmoninarum infections. We challenged Chinook salmon with R. salmoninarum at 12 °C for 2 weeks and then divided the fish into three temperature groups (8, 12 and 15 °C). Fish in the 8 °C group had significantly higher R. salmoninarum‐specific mortality, kidney R. salmoninarum loads and bacterial shedding rates relative to the fish held at 12 or 15 °C. There was a trend towards suppressed bacterial load and shedding in the 15 °C group, but the results were not significant. Bacterial load was a significant predictor of shedding for the 8 and 12 °C groups but not for the 15 °C group. Overall, our results showed little effect of temperature stress on the progress of infection, but do support the conclusion that cooler water temperatures contribute to infection progression and increased transmission potential in Chinook salmon infected with R. salmoninarum.     

Survival of smaller sport caught chinook, Oncorhynchus tshawytscha (Walbaum), and coho, Oncorhynchus kisutch (Walbaum), salmon from Lake Michigan and its management implications

Abstract The survival of small-sized (<50.8 cm) chinook salmon, Oncorhynchus tshawytscha (Walbaum), and coho salmon, Oncorhynchus kitsutch (Walbaum), caught by sport fishing was determined to assess the potential for increasing the size limit for these fish. Fishermen were recently catching smaller salmon than in the 1970s, but salmon growth rates had not changed. To be an effective management option, the survival rate of hooked and returned fish must be high. The overall survival rates were high: 76% for chinook salmon and 70% for coho salmon. There was no significant difference in survival of the coho salmon with size of hook used ( P = 0.31). Any mortality among fish was generally acute; fish hooked deep in the mouth or gills generally bled and died shortly after capture. Fish hooked in the gills had a significantly greater mortality ( P = 0.0002). The overall high survival rate for these species was the result of a small proportion of fish being hooked in the gills or deep in the mouth. Since the survival rate of the salmon was high, the size limit could be increased to allow smaller fish to grow to sizes preferred by sport fishermen.     

Evaluation of Juvenile salmon passage and survival through a fish weir and other routes at Foster Dam,Oregon, USA

Downstream passage for juvenile Chinook salmon Oncorhynchus tshawytscha (Walbaum) and steelhead Oncorhynchus mykiss (Walbaum) through the fish weir at Foster Dam, Oregon, was evaluated to assess the weir's efficacy as a long-term passage solution. Radio telemetry was used to estimate survival, passage and effectiveness for the fish weir, spillway and turbines. Survival of Chinook salmon through the fish weir varied by season and reservoir elevation (66%–87%), and passage proportions were low (8%–20%). Fish weir effectiveness was low to moderate (0.39–2.09). Steelhead survival was lower (57%–77%) but passed the fish weir in higher proportions (77%), and weir effectiveness was consistently high (2.96–5.49). The results indicate the existing fish weir will not be a suitable long-term passage solution for all species. However, because of the high passage proportions and effectiveness observed for steelhead, further development of the surface-flow concept appears to be promising for safely passing downstream-migrating fishes.     

Bacterial infections of Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to gamete collecting weirs in Michigan

Herein, we describe the prevalence of bacterial infections in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to spawn in two tributaries within the Lake Michigan watershed. Ten bacterial genera, including Renibacterium, Aeromonas, Carnobacterium, Serratia, Proteus, Pseudomonas, Hafnia, Salmonella, Shewanella and Morganella, were detected in the kidneys of Chinook salmon (n = 480) using culture, serological and molecular analyses. Among these, Aeromonas salmonicida was detected at a prevalence of ～15%. Analyses revealed significant interactions between location/time of collection and gender for these infections, whereby overall infection prevalence increased greatly later in the spawning run and was significantly higher in females. Renibacterium salmoninarum was detected in fish kidneys at an overall prevalence of >25%. Logistic regression analyses revealed that R. salmoninarum prevalence differed significantly by location/time of collection and gender, with a higher likelihood of infection later in the spawning season and in females vs. males. Chi‐square analyses quantifying non‐independence of infection by multiple pathogens revealed a significant association between R. salmoninarum and motile aeromonad infections. Additionally, greater numbers of fish were found to be co‐infected by multiple bacterial species than would be expected by chance alone. The findings of this study suggest a potential synergism between bacteria infecting spawning Chinook salmon.     

Effects of temperature on disease progression and swimming stamina in Ichthyophonus-infected rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss , were infected with Ichthyophonus sp. and held at 10 °C, 15 °C and 20 °C for 28 days to monitor mortality and disease progression. Infected fish demonstrated more rapid onset of disease, higher parasite load, more severe host tissue reaction and reduced mean-day-to-death at higher temperature. In a second experiment, Ichthyophonus -infected fish were reared at 15 °C for 16 weeks then subjected to forced swimming at 10 °C, 15 °C and 20 °C. Stamina improved significantly with increased temperature in uninfected fish; however, this was not observed for infected fish. The difference in performance between infected and uninfected fish became significant at 15 °C ( P  =   0.02) and highly significant at 20 °C ( P  =   0.005). These results have implications for changes in the ecology of fish diseases in the face of global warming and demonstrate the effects of higher temperature on the progression and severity of ichthyophoniasis as well as on swimming stamina, a critical fitness trait of salmonids. This study helps explain field observations showing the recent emergence of clinical ichthyophoniasis in Yukon River Chinook salmon later in their spawning migration when water temperatures were high, as well as the apparent failure of a substantial percentage of infected fish to successfully reach their natal spawning areas.     

Spatial distribution of ocean habitat of yearling Chinook (Oncorhynchus tshawytscha) and coho (Oncorhynchus kisutch) salmon off Washington and Oregon,USA

We determined the habitat usage and habitat connectivity of juvenile Chinook (Oncorhynchus tshawytscha) and coho (Oncorhynchus kisutch) salmon in continental shelf waters off Washington and Oregon, based on samples collected every June for 9 yr (1998–2006). Habitat usage and connectivity were evaluated using SeaWiFS satellite‐derived chlorophyll a data and water depth. Logistic regression models were developed for both species, and habitats were first classified using a threshold value estimated from a receiver operating characteristic curve. A Bernoulli random process using catch probabilities from observed data, i.e. the frequency of occurrence of a fish divided by the number of times a station was surveyed, was applied to reclassify stations. Zero‐catch probabilities of yearling Chinook and yearling coho salmon decreased with increases in chlorophyll a concentration, and with decreases in water depth. From 1998 to 2006, ～ 47% of stations surveyed were classified as unfavorable habitat for yearling Chinook salmon and ～ 53% for yearling coho salmon. Potentially favorable habitat varied among years and ranged from 9 856 to 15 120 km² (Chinook) and from 14 800 to 16 736 km² (coho). For both species, the smallest habitat area occurred in 1998, an El Niño year. Favorable habitats for yearling Chinook salmon were more isolated in 1998 and 2005 than in other years. Both species had larger and more continuous favorable habitat areas along the Washington coast than along the Oregon coast. The favorable habitats were also larger and more continuous nearshore than offshore for both species. Further investigations on large‐scale transport, mesoscale physical features, and prey and predator availability in the study area are necessary to explain the spatial arrangement of juvenile salmon habitats in continental shelf waters.     

Evaluation of an experimental Aeromonas salmonicida epidemic in chinook salmon, Oncorhynchus tshawytscha (Walbaum)

To determine the dynamics of the transmission of Aeromonas salmonicida ssp. salmonicida infection, chinook salmon, Oncorhynchus tshawytscha, were exposed to bacteria by cohabitation. The latent period (time between exposure and infectivity) was determined by exposing a group of chinook salmonid fingerlings to A. salmonicida by bath, then, at daily intervals, by holding five exposed (donor) fish with approximately 50 naive fish for 24 h. The latent period was 3 days post-infection and the time period between the initial exposure to bacteria and the beginning of bacterial shedding was 4.5 days for the same animals. The prevalence and intensity of infection in the donor fish, to which recipient fish were exposed, i.e. the level of exposure, was highly correlated with the development of disease in recipient (susceptible) chinook salmon (r2 = 0.57). An experiment was conducted to determine the daily progress of infection and development of a furunculosis epidemic among recipient fish by cohabiting a single exposed fish with 43 unexposed salmon. At daily intervals, all fish (in seven treatment tanks and one control tank daily) were sacrificed and tested for the presence of A. salmonicida in the kidney (n = 3520). Over 10 days, mean prevalence among recipient fish reached 75% and disease related mortality exceeded 50%. Bacterial concentrations in the water continued to increase over the duration of the experiment in concert with the number of infected animals present in the population.     

Immunosuppression in rainbow trout, Salmo gairdneri Richardson, caused by the haemoflagellate Cryptobia salmositica Katz, 1951

Abstract. An experimental Cryptobia salmositica infection in rainbow trout, Salmo gairdneri Richardson, produced suppression of the humoral response against sheep red blood cells as measured by direct haemagglutination. Two-month and 5-month infections produced equal suppression. The parasite also produced suppression of the humoral response against a bacterial pathogen, Yersinia ruckeri . Anti- Y. ritckeri titres were significantly lower in most fish infected with C. salmositica than in non-infected fish. Immunosuppression became evident when C. salmositica first appeared in the blood (first 2 weeks of infection), Immunosuppression was confirmed by challenge with Y. ruckeri . Mortality at challenge occurred in 64·3% to 83·3% of the fish already infected with C. salmositica at the time of initial Y. ruckeri exposure. There was no mortality at challenge if fish were not infected with C. salmositica at initial bacterial exposure, nor in those concurrently infected with both pathogens. Antigenic competition may have caused the immunosuppression.     

Piscine reovirus,but not Jaundice Syndrome,was transmissible to Chinook Salmon,Oncorhynchus tshawytscha (Walbaum), Sockeye Salmon,Oncorhynchus nerka (Walbaum), and Atlantic Salmon,Salmo salar L.

A Jaundice Syndrome occurs sporadically among sea‐pen‐farmed Chinook Salmon in British Columbia, the westernmost province of Canada. Affected salmon are easily identified by a distinctive yellow discolouration of the abdominal and periorbital regions. Through traditional diagnostics, no bacterial or viral agents were cultured from tissues of jaundiced Chinook Salmon; however, piscine reovirus (PRV) was identified via RT‐rPCR in all 10 affected fish sampled. By histopathology, Jaundice Syndrome is an acute to peracute systemic disease, and the time from first clinical signs to death is likely <48 h; renal tubular epithelial cell necrosis is the most consistent lesion. In an infectivity trial, Chinook Salmon, Sockeye Salmon and Atlantic Salmon, intraperitoneally inoculated with a PRV‐positive organ homogenate from jaundiced Chinook Salmon, developed no gross or microscopic evidence of jaundice despite persistence of PRV for the 5‐month holding period. The results from this study demonstrate that the Jaundice Syndrome was not transmissible by injection of material from infected fish and that PRV was not the sole aetiological factor for the condition. Additionally, these findings showed the Pacific coast strain of PRV, while transmissible, was of low pathogenicity for Atlantic Salmon, Chinook Salmon and Sockeye Salmon.     

Development and validation of real-time PCR for the detection of Yersinia ruckeri

Yersiniosis (enteric red mouth disease) is a contagious bacterial disease caused by Yersinia ruckeri, which primarily affects salmonids. A real-time PCR assay using a molecular beacon has been developed and validated to improve the detection of the causative biotypes of Y. ruckeri. The assay, which targets the glnA (glutamine synthetase) gene, proved to have 100% analytical specificity and analytical sensitivities of 5 fg and 3 × 10(3) CFU g(-1) for DNA and seeded kidney tissue, respectively. The assay was highly repeatable with low % CV for intra- and inter-run experiments, and the optimized parameters transferred easily between different real-time PCR platforms. Following analytical validation, diagnostic specificity was determined using New Zealand farmed Chinook salmon (n = 750) from 10 farms during 2007/08. The real-time PCR was run in parallel with the bacterial culture detection method, and all fish tested were found to be negative by both methods for Y. ruckeri, resulting in 100% diagnostic specificity (95% confidence interval). The molecular beacon real-time PCR system is specific, sensitive, reproducible and a rapid method for the detection of Y. ruckeri and has the potential to be used for routine diagnostic testing, health certification and active surveillance programmes.     

Siderophores produced by Vibrio anguillarum in vitro and in infected rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. Ten strains of Vibrio anguillarum produced three different types of iron-binding compounds when cultured under different conditions. These were (1) a common phenolate siderophore produced by all strains. (2) a hydroxamate siderophore produced by three strains and (3) a second phenolate siderophore, tentatively identified as anguibactin, produced by V. anguillarum strain 775 and two other strains, all of which contained a plasmid of 45–50 Md. The relative affinities of these siderophores, determined by competition for ⁵⁵Fe was: anguibactin < hydroxamate siderophore < common phenolate siderophore. However, under these conditions, none removed iron from purified aerobactin. Experimental infection of rainbow trout. Oncorhynchus mykiss (Walbaum), showed that only the common phenolate siderophore was detected in the kidney and spleen of fish infected with strains 91079 and NCIMB6. The hydroxamate siderophore produced in vitro by strain NCIMB6 was not detected in vivo. However, in the kidney of fish infected with strain 775, both the common phenolate siderophore and anguibactin were detected, showing that a second uptake system is required by strain 775 in vivo and that the iron-uptake system based on the common phenolate siderophore is defective.     

Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum)

The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management.